Autolysis of Pichia pastoris induced by cold View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2017-05-12

AUTHORS

Yaneth Bartolo-Aguilar, Luc Dendooven, Cipriano Chávez-Cabrera, Luis B. Flores-Cotera, María E. Hidalgo-Lara, Lourdes Villa-Tanaca, Rodolfo Marsch

ABSTRACT

The production of recombinant biopharmaceutical proteins is a multi-billion dollar market. Protein recovery represents a major part of the production costs. Pichia pastoris is one of the microbial systems most used for the production of heterologous proteins. The use of a cold-induced promoter to express lytic enzymes in the yeast after the growth stage could reduce protein recovery costs. This study shows that a cold-shock can be applied to induce lysis of the yeast cells. A strain of P. pastoris was constructed in which the endogenous eng gene encoding a putative endo-β-1,3-glucanase was overexpressed using the cold-shock induced promoter of the cctα gene from Saccharomyces cerevisiae. In the transgenic P. pastoris, the expression of eng increased 3.6-fold after chilling the cells from 30 to 4 °C (cold-shock stage) followed by incubation for 6 h (eng expression stage). The culture was heated to 30 °C for 6 h (ENG synthesis stage) and kept at 37 °C for 24 h (lysis stage). After this procedure the cell morphology changed, spheroplasts were obtained and cellular lysis was observed. Thus, a clone of P. pastoris was obtained, which undergoes autolysis after a cold-shock. More... »

PAGES

95

Identifiers

URI

http://scigraph.springernature.com/pub.10.1186/s13568-017-0397-y

DOI

http://dx.doi.org/10.1186/s13568-017-0397-y

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1085379512

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/28500590


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